CN106857205A - The efficient authentication method of alfalfa variety drought-resistant ability - Google Patents
The efficient authentication method of alfalfa variety drought-resistant ability Download PDFInfo
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Abstract
本发明公开了苜蓿品种抗旱能力的高效鉴定方法，包括以下步骤：进行聚乙二醇PEG‑6000模拟干旱胁迫处理；测定不同抗旱级别苜蓿品种响应不同胁迫强度时变化最敏感的生理指标及其敏感阈值；建立与苜蓿抗旱级别相对应的敏感性生理指标群及敏感阈值指标体系；将待鉴定的苜蓿品种划分为抗旱性强、中、弱3 类。本发明提供的鉴定方法，采用不同浓度PEG‑6000模拟干旱胁迫时，依据与不同抗旱级别苜蓿密切相关的生理指标及其敏感阈值作为鉴定依据，建立与抗旱级别相应的敏感性生理指标群及敏感阈值指标体系，对苜蓿抗旱能力进行分级，提高苜蓿抗旱性综合评价结果的准确性，具有很好的操作性和可靠性，亦可在实验室完成抗旱能力的鉴定，条件易于控制，成本低。The invention discloses a high-efficiency identification method for the drought resistance ability of alfalfa varieties, which comprises the following steps: carrying out polyethylene glycol PEG-6000 simulated drought stress treatment; measuring the most sensitive physiological index and its sensitivity of alfalfa varieties with different drought resistance levels in response to different stress intensities. Threshold; establish a sensitive physiological index group and a sensitive threshold index system corresponding to the drought resistance level of alfalfa; divide the alfalfa varieties to be identified into three categories of strong, medium and weak drought resistance. In the identification method provided by the present invention, when using different concentrations of PEG-6000 to simulate drought stress, based on the physiological indicators closely related to different drought-resistant levels of alfalfa and their sensitive thresholds as the identification basis, the sensitive physiological index groups and sensitive physiological indicators corresponding to the drought-resistant levels are established. The threshold index system can grade alfalfa drought resistance and improve the accuracy of comprehensive evaluation results of alfalfa drought resistance. It has good operability and reliability, and can also complete the identification of drought resistance in the laboratory. The conditions are easy to control and the cost is low.
Description
技术领域technical field
本发明涉及植物抗旱性鉴定技术领域，具体涉及苜蓿品种抗旱能力的高效鉴定方法。The invention relates to the technical field of plant drought resistance identification, in particular to an efficient identification method for the drought resistance ability of alfalfa varieties.
背景技术Background technique
苜蓿抗旱性鉴定常伴随苜蓿引种与选育进行。通常把抗旱性不同的苜蓿品种在田间栽培或室内培养，在正常水分条件(对照)或胁迫强度增大的干旱胁迫下，使其幼苗生长或处理一定的时间，比较各苜蓿品种受不同干旱胁迫影响后其形态和生理指标的变化，通过二者的相关分析，对其抗旱性作综合评定。在紫花苜蓿的抗旱性鉴定中，重点在于抗旱性指标的选择，归纳起来有形态指标、生理指标、生长发育指标、生产性能指标等四大类。一般包括根、叶形态指标、相对含水量、叶水势、质膜透性、 脯氨酸含量、酶活力、根冠比、产量等。也可先利用限制性片段长度多态性(RFIP)甄别抗旱基因的存在并进行抗旱基因定位，建立RELP遗传连锁来鉴定其抗旱性。目前较常见的多以田间指标或者结合若干生理指标对紫花苜蓿的抗旱性作出评价，但与抗旱性相关的生理生化指标有许多，随着研究的进一步深入，人们对于某些指标与抗旱性的关系产生了疑问，某些指标与抗旱性并非呈简单的线性关系，且不同胁迫程度下，与苜蓿抗旱性密切相关的指标也不确定。由于苜蓿的抗旱性受多因素的影响，适当的鉴定指标是准确评价苜蓿抗旱性的关键。因此，简单、可靠的抗旱鉴定方法和准确高效的鉴定指标的正确选择在苜蓿抗旱能力鉴定中具有重要意义。The drought resistance identification of alfalfa is often accompanied by the introduction and breeding of alfalfa. Usually, alfalfa varieties with different drought resistance are cultivated in the field or indoors, and the seedlings are grown or treated for a certain period of time under normal water conditions (control) or drought stress with increased stress intensity. The changes of its morphology and physiological indexes after the impact, through the correlation analysis of the two, make a comprehensive evaluation of its drought resistance. In the identification of drought resistance of alfalfa, the focus is on the selection of drought resistance indicators, which can be summarized into four categories: morphological indicators, physiological indicators, growth and development indicators, and production performance indicators. Generally include root and leaf morphological indicators, relative water content, leaf water potential, plasma membrane permeability, proline content, enzyme activity, root-to-shoot ratio, yield, etc. Restriction fragment length polymorphism (RFIP) can also be used to identify the existence of drought-resistant genes and map the drought-resistant genes, and establish RELP genetic linkage to identify their drought-resistant properties. At present, it is more common to evaluate the drought resistance of alfalfa with field indicators or a combination of several physiological indicators, but there are many physiological and biochemical indicators related to drought resistance. The relationship between some indicators and drought resistance was not a simple linear relationship, and under different stress levels, the indicators closely related to drought resistance of alfalfa were also uncertain. Since the drought resistance of alfalfa is affected by many factors, appropriate identification indexes are the key to accurately evaluate the drought resistance of alfalfa. Therefore, a simple and reliable drought resistance identification method and the correct selection of accurate and efficient identification indicators are of great significance in the identification of alfalfa drought resistance.
现有技术中，有关苜蓿抗旱性鉴定的技术较多，主要是采用温室盆栽模拟旱境或不同水势PEG-6000模拟干旱胁迫结合测定若干生理指标的方法鉴定不同苜蓿品种的抗旱性能，并采用相应的评定方法，如直接比较法、总抗旱性评价法、隶属函数法、层次分析法(AHP)、主成分分析法和灰色关联度模型综合评价苜蓿品种的抗旱能力。李慥哲（1991）用聚乙二醇 (PEG-6000) 溶液模拟干旱胁迫条件，测定了10个苜蓿品种组织相对含水量、地上部干物质产量、相对生长率、细胞膜透性、过氧化物酶活性、游离脯氨酸含量、叶片持水力、高渗溶液下种子的发芽率、反复干旱幼苗成活率。应用主成分分析对这些苜蓿品种幼苗的抗旱性进行了综合评定，使其归属于强抗旱、中抗旱和弱抗旱3 个抗旱等级。康俊梅等（2004）对来自国内外不同种质的 41 个紫花苜蓿品种进行盆栽试验，测定了水分饱和亏缺、死亡率、干物质胁迫指数、根系胁迫指数、地上生物量、地下生物量等7个抗旱鉴定指标，应用系统聚类的方法对数据进行分级聚类，使其归属于强抗旱、中抗旱和弱抗旱3 个抗旱等级。孟林等（2008）对17 个苜蓿品种苗期干旱后的叶片相对含水量、脯氨酸含量和质膜透性 3 个抗旱生理指标进行了分析评价，并将17 个苜蓿品种划分为抗旱性较强、中等、较弱3 类。In the prior art, there are many technologies related to the identification of drought resistance of alfalfa. The main method is to identify the drought resistance performance of different alfalfa varieties by using the simulated drought environment of greenhouse pot planting or the simulated drought stress of different water potential PEG-6000 combined with the determination of several physiological indicators, and using corresponding Evaluation methods, such as direct comparison method, total drought resistance evaluation method, subordination function method, analytic hierarchy process (AHP), principal component analysis method and gray relational degree model to comprehensively evaluate the drought resistance ability of alfalfa varieties. Li Zhenzhe (1991) used polyethylene glycol (PEG-6000) solution to simulate drought stress conditions, and measured tissue relative water content, shoot dry matter production, relative growth rate, cell membrane permeability, and peroxidase activity of 10 alfalfa varieties. , free proline content, water holding capacity of leaves, germination rate of seeds under hypertonic solution, survival rate of seedlings after repeated drought. The drought resistance of the seedlings of these alfalfa varieties was comprehensively evaluated by principal component analysis, and they belonged to three drought resistance grades: strong drought resistance, medium drought resistance and weak drought resistance. Kang Junmei et al. (2004) conducted pot experiments on 41 alfalfa varieties from different germplasms at home and abroad, and measured water saturation deficit, mortality, dry matter stress index, root stress index, aboveground biomass, underground biomass, etc. The 7 drought resistance identification indexes were classified into 3 drought resistance grades: strong drought resistance, medium drought resistance and weak drought resistance. Meng Lin et al. (2008) analyzed and evaluated the three drought-resistant physiological indicators of 17 alfalfa varieties after seedling drought: relative leaf water content, proline content and plasma membrane permeability, and classified 17 alfalfa varieties into drought-resistant There are 3 categories: Strong, Medium, and Weak.
以上采用温室盆栽模拟旱境或PEG模拟干旱结合测定抗旱生理指标评价苜蓿幼苗抗旱能力时，其评价方法均为统一指标体系下的综合评价，仅考虑不同抗旱级别苜蓿的相同抗旱生理指标类别对干旱胁迫的敏感性存在差异，但未考虑不同胁迫强度下不同抗旱级别苜蓿的敏感性生理指标类别群存在差异性的特征，故选择的抗旱生理指标缺乏特殊性和代表性，进而影响了苜蓿抗旱性综合评价结果的准确度。When using greenhouse pot plant simulated drought environment or PEG simulated drought combined with determination of drought resistance physiological indicators to evaluate the drought resistance ability of alfalfa seedlings, the evaluation method is the comprehensive evaluation under the unified index system, only considering the effect of the same drought resistance physiological index category of alfalfa with different drought resistance levels on drought There are differences in the sensitivity to stress, but the differences in the sensitivity physiological indicators of alfalfa with different drought resistance levels under different stress intensities are not considered. Therefore, the selected drought resistance physiological indicators lack specificity and representativeness, which in turn affects the drought resistance of alfalfa. Comprehensive evaluation of the accuracy of the results.
发明内容Contents of the invention
本发明的目的就是针对上述现有技术中的缺陷，提供了一种苜蓿品种抗旱能力的高效鉴定方法，是采用不同浓度PEG-6000模拟干旱胁迫，分别测定不同抗旱胁迫程度下苜蓿品种最敏感的生理指标；并根据各胁迫程度下敏感性生理指标类别群差异特征，将苜蓿品种划分为强抗旱、中抗旱和弱抗旱苜蓿，以便简单、准确高效地鉴定苜蓿品种的抗旱能力。The object of the present invention is to aim at the defects in the above-mentioned prior art, and to provide a kind of high-efficiency identification method of alfalfa variety drought resistance, which is to adopt different concentrations of PEG-6000 to simulate drought stress, and measure the most sensitive alfalfa variety under different levels of drought resistance stress respectively. Physiological indicators; and according to the difference characteristics of sensitive physiological indicators under various stress levels, alfalfa varieties were divided into strong drought-resistant, medium drought-resistant and weak drought-resistant alfalfa, so as to identify the drought resistance of alfalfa varieties simply, accurately and efficiently.
为了实现上述目的，本发明提供的技术方案为：苜蓿品种抗旱能力的高效鉴定方法，包括以下步骤：In order to achieve the above object, the technical solution provided by the present invention is: an efficient identification method for drought resistance of alfalfa varieties, comprising the following steps:
1）进行聚乙二醇PEG-6000模拟干旱胁迫处理：在室温25℃下，用等量含不同浓度聚乙二醇PEG-6000的Hoagland’s营养液根际胁迫苜蓿幼苗7d；1) Simulated drought stress treatment with polyethylene glycol PEG-6000: at room temperature 25°C, the rhizosphere stress of alfalfa seedlings with equal amounts of Hoagland’s nutrient solution containing different concentrations of polyethylene glycol PEG-6000 for 7 days;
2）测定不同胁迫强度下，不同抗旱级别苜蓿品种响应各胁迫强度时变化最敏感的生理指标及其敏感阈值；2) Under different stress intensities, the most sensitive physiological indicators and their sensitive thresholds of alfalfa varieties with different drought resistance levels in response to various stress intensities were determined;
3）选取与不同抗旱级别苜蓿密切相关的生理指标及其敏感阈值作为鉴定依据，建立与苜蓿抗旱级别相对应的敏感性生理指标群及敏感阈值指标体系；3) Select the physiological indicators closely related to different drought-resistant levels of alfalfa and their sensitive thresholds as the identification basis, and establish a sensitive physiological index group and sensitive threshold index system corresponding to the drought-resistant levels of alfalfa;
4）根据建立的与苜蓿抗旱级别相对应的敏感性生理指标群及敏感阈值指标体系，将待鉴定的苜蓿品种划分为抗旱性强、中、弱3 类。4) According to the established sensitive physiological index group and sensitive threshold index system corresponding to the drought resistance level of alfalfa, the alfalfa varieties to be identified were divided into three categories: strong, medium and weak drought resistance.
进一步的，上述的苜蓿品种抗旱能力的高效鉴定方法，所述步骤2）中，测定得到不同抗旱级别苜蓿对应的生理指标群所包含的最敏感的生理指标分别为：Further, in the above-mentioned efficient identification method for the drought resistance ability of alfalfa varieties, in the step 2), the most sensitive physiological indicators included in the physiological index groups corresponding to different drought resistance levels of alfalfa are determined as follows:
a）0-0.18g/mL的PEG-6000轻度胁迫下，最敏感的生理指标为脯氨酸含量、可溶性蛋白含量、过氧化氢酶活性、过氧化氢含量和羟自由基浓度；a) Under mild stress of 0-0.18g/mL PEG-6000, the most sensitive physiological indicators are proline content, soluble protein content, catalase activity, hydrogen peroxide content and hydroxyl radical concentration;
b）0.18-0.33 g/mL的PEG-6000中度胁迫下，最敏感的生理指标为过氧化氢含量、叶片相对含水量、还原性抗坏血酸含量和SOD活性；b) Under moderate stress of 0.18-0.33 g/mL PEG-6000, the most sensitive physiological indicators were hydrogen peroxide content, relative leaf water content, reduced ascorbic acid content and SOD activity;
c）0.33-0.43g/mL的PEG-6000重度胁迫下，最敏感的生理指标为还原性抗坏血酸含量、丙二醛含量、还原性谷胱甘肽含量、过氧化氢酶活性、超氧阴离子自由基含量和叶绿素含量。c) Under severe stress of 0.33-0.43g/mL PEG-6000, the most sensitive physiological indicators are reduced ascorbic acid content, malondialdehyde content, reduced glutathione content, catalase activity, superoxide anion free basal content and chlorophyll content.
进一步的，上述的苜蓿品种抗旱能力的高效鉴定方法，所述步骤3）中，最敏感的生理指标阈值及其构建的划分标准体系指标的具体划分方式为：Further, in the above-mentioned efficient identification method of drought resistance of alfalfa varieties, in the step 3), the specific division method of the most sensitive physiological index threshold and the index of the classification standard system constructed is as follows:
A）0-0.18g/mL的轻度胁迫，强抗旱苜蓿过氧化氢和羟自由基含量变化最敏感，且分别低于32.12μmol·g-1和1891.22 A·1000·g-1；中抗旱苜蓿过氧化氢酶活性变化最敏感，且介于46.59-75.47U·g-1·min-1；弱抗旱苜蓿脯氨酸和可溶性蛋白含量变化最敏感，且脯氨酸含量低于288.26µg·g-1、可溶性蛋白含量高于6.42 mg.g-1；A) Mild stress of 0-0.18g/mL, strong drought-resistant alfalfa hydrogen peroxide and hydroxyl radical content changes are the most sensitive, and they are lower than 32.12μmol·g -1 and 1891.22 A·1000·g -1 respectively; moderate drought resistance The change of catalase activity in alfalfa was the most sensitive, and it was between 46.59-75.47U·g -1 ·min -1 ; the content of proline and soluble protein in weakly drought-resistant alfalfa was the most sensitive, and the content of proline was lower than 288.26µg· g -1 , soluble protein content higher than 6.42 mg.g -1 ;
B）0.18-0.33 g/mL的中度胁迫，强抗旱苜蓿叶片相对含水量和过氧化氢含量变化最敏感，且叶片相对含水量大于50.43%、过氧化氢含量低于78.27μmol·g-1；中抗旱苜蓿还原性抗坏血酸含量变化最敏感 ，且介于60.44-215.10μg·g-1 ；弱抗旱苜蓿叶片相对含水量和SOD活性变化最敏感，且叶片相对含水量小于32.06%、SOD活性低于360.64 U·g-1；B) Under moderate stress of 0.18-0.33 g/mL, the leaves of strong drought-resistant alfalfa are most sensitive to changes in relative water content and hydrogen peroxide content, and the relative water content of leaves is greater than 50.43%, and the content of hydrogen peroxide is lower than 78.27 μmol·g -1 ; The content of reducing ascorbic acid in medium drought-resistant alfalfa is the most sensitive, and it is between 60.44-215.10 μg·g -1 ; The change of relative water content and SOD activity in leaves of weak drought-resistant alfalfa is the most sensitive, and the relative water content of leaves is less than 32.06%, and the SOD activity is low at 360.64 U·g -1 ;
C）0.33-0.43g/mL的重度胁迫，强抗旱苜蓿还原性抗坏血酸和还原性谷胱甘肽含量变化最敏感，且分别高于432.95μg·g-1和5280.17μmol·g-1；中抗旱苜蓿丙二醛含量变化最敏感，且介于 0.13-0.15μmol·g-1；弱抗旱苜蓿过氧化氢酶活性、还原性谷胱甘肽含量、叶绿素含量和超氧阴离子自由基含量变化最敏感，且过氧化氢酶活性低于297.45 U·g-1·min-1、还原性谷胱甘肽含量低于2394.42μmol·g-1、叶绿素含量低于2.46 mg.g-1、超氧阴离子自由基含量高于152.92 nmol·g-1·min-1；C) Under severe stress of 0.33-0.43g/mL, the contents of reduced ascorbic acid and reduced glutathione in alfalfa with strong drought resistance were the most sensitive, and they were higher than 432.95μg·g -1 and 5280.17μmol·g -1 respectively; moderate drought resistance The change of malondialdehyde content in alfalfa is the most sensitive, and it is between 0.13-0.15μmol·g -1 ; the change of catalase activity, reduced glutathione content, chlorophyll content and superoxide anion free radical content in weak drought-resistant alfalfa is the most sensitive , and the catalase activity is lower than 297.45 U·g -1 ·min -1 , the reduced glutathione content is lower than 2394.42 μmol·g -1 , the chlorophyll content is lower than 2.46 mg.g -1 , superoxide anion The free radical content is higher than 152.92 nmol·g -1 ·min -1 ;
D）同时满足所述A）、B）、C）中，强抗旱苜蓿指标群的各指标阈值要求，则归为强抗旱苜蓿；同时满足所述A）、B）、C）中，中抗旱苜蓿指标群的各指标阈值要求，则归为中抗旱苜蓿；同时满足所述A）、B）、C）中，弱抗旱苜蓿指标群的各指标阈值要求，则归为弱抗旱苜蓿品种。D) Satisfying the threshold requirements of each indicator group of strong drought-resistant alfalfa in A), B), and C) at the same time, it is classified as strong drought-resistant alfalfa; meeting the above-mentioned A), B), and C) at the same time, moderate drought resistance Alfalfa varieties that meet the threshold requirements for each index of the alfalfa index group are classified as medium drought-resistant alfalfa varieties; those that meet the threshold requirements for each indicator group of weak drought-resistant alfalfa in A), B), and C) are classified as weak drought-resistant alfalfa varieties.
本发明的有益效果为：本发明提供的苜蓿品种抗旱能力的高效鉴定方法，在采用不同浓度PEG-6000模拟干旱胁迫时，在考虑不同抗旱级别苜蓿品种敏感性生理指标类别体系存在差异的基础上，选取不同胁迫程度下与苜蓿抗旱性密切相关的生理指标作为鉴定指标，根据不同胁迫强度相应的敏感性生理指标群及其指标敏感阈值对苜蓿的抗旱能力进行分级，提高苜蓿抗旱性综合评价结果的准确性，从而准确地鉴定苜蓿品种的抗旱能力，具有很好的实用性和可靠性。此外，本发明的方法可在实验室完成苜蓿抗旱能力的鉴定，试验条件易于控制，成本较低，操作性强。The beneficial effects of the present invention are as follows: the high-efficiency identification method for the drought resistance ability of alfalfa varieties provided by the present invention, when using different concentrations of PEG-6000 to simulate drought stress, on the basis of considering the differences in the sensitive physiological index category systems of alfalfa varieties with different levels of drought resistance Select the physiological indicators closely related to the drought resistance of alfalfa under different stress levels as identification indicators, and classify the drought resistance of alfalfa according to the corresponding sensitive physiological index groups and index sensitivity thresholds of different stress intensities, and improve the comprehensive evaluation results of alfalfa drought resistance. The accuracy, so as to accurately identify the drought resistance of alfalfa varieties, has good practicability and reliability. In addition, the method of the invention can complete the identification of alfalfa drought resistance in a laboratory, the test conditions are easy to control, the cost is low, and the operability is strong.
具体实施方式detailed description
实施例1：Example 1:
苜蓿品种抗旱能力的高效鉴定方法，包括以下步骤：An efficient identification method for drought resistance of alfalfa varieties, comprising the following steps:
1）进行聚乙二醇PEG-6000模拟干旱胁迫处理：在室温25℃下，用等量含不同浓度聚乙二醇PEG-6000的Hoagland’s营养液根际胁迫苜蓿幼苗7d；1) Simulated drought stress treatment with polyethylene glycol PEG-6000: at room temperature 25°C, the rhizosphere stress of alfalfa seedlings with equal amounts of Hoagland’s nutrient solution containing different concentrations of polyethylene glycol PEG-6000 for 7 days;
2）测定不同胁迫强度下，不同抗旱级别苜蓿品种响应各胁迫强度时变化最敏感的生理指标；测定得到的最敏感的生理指标分别为：2) Under different stress intensities, the most sensitive physiological indicators of alfalfa varieties with different drought resistance levels in response to the stress intensities were measured; the most sensitive physiological indicators obtained were:
a）0-0.18g/mL的轻度胁迫下，最敏感的生理指标为脯氨酸含量、可溶性蛋白含量、过氧化氢酶活性、过氧化氢含量和羟自由基浓度；a) Under mild stress of 0-0.18g/mL, the most sensitive physiological indicators are proline content, soluble protein content, catalase activity, hydrogen peroxide content and hydroxyl radical concentration;
b）0.18-0.33 g/mL的中度胁迫下，最敏感的生理指标为过氧化氢含量、叶片相对含水量、还原性抗坏血酸含量和SOD活性；b) Under moderate stress of 0.18-0.33 g/mL, the most sensitive physiological indicators were hydrogen peroxide content, relative leaf water content, reduced ascorbic acid content and SOD activity;
c）0.33-0.43g/mL的重度胁迫下，最敏感的生理指标为还原性抗坏血酸含量、还原性谷胱甘肽含量、过氧化氢酶活性、超氧阴离子自由基含量和叶绿素含量；c) Under severe stress of 0.33-0.43g/mL, the most sensitive physiological indicators are reduced ascorbic acid content, reduced glutathione content, catalase activity, superoxide anion free radical content and chlorophyll content;
3）根据各生理指标对干旱胁迫的敏感阈值，将待鉴定的苜蓿品种划分为抗旱性强、中、弱3 类，划分的具体方式为：3) According to the sensitivity threshold of each physiological index to drought stress, the alfalfa varieties to be identified are divided into three categories: strong, medium and weak in drought resistance. The specific method of division is as follows:
A）0-0.18g/mL的轻度胁迫，强抗旱苜蓿过氧化氢和羟自由基含量变化最敏感，且分别低于32.12μmol·g-1和1891.22 A·1000·g-1；中抗旱苜蓿过氧化氢酶活性变化最敏感，且介于46.59-75.47U·g-1·min-1；弱抗旱苜蓿脯氨酸和可溶性蛋白含量变化最敏感，且脯氨酸含量低于288.26µg·g-1、可溶性蛋白含量高于6.42 mg.g-1；A) Mild stress of 0-0.18g/mL, strong drought-resistant alfalfa hydrogen peroxide and hydroxyl radical content changes are the most sensitive, and they are lower than 32.12μmol·g -1 and 1891.22 A·1000·g -1 respectively; moderate drought resistance The change of catalase activity in alfalfa was the most sensitive, and it was between 46.59-75.47U·g -1 ·min -1 ; the content of proline and soluble protein in weakly drought-resistant alfalfa was the most sensitive, and the content of proline was lower than 288.26µg· g -1 , soluble protein content higher than 6.42 mg.g -1 ;
B）0.18-0.33 g/mL的中度胁迫，强抗旱苜蓿叶片相对含水量和过氧化氢含量变化最敏感，且叶片相对含水量大于50.43%、过氧化氢含量低于78.27μmol·g-1；中抗旱苜蓿还原性抗坏血酸含量变化最敏感 ，且介于60.44-215.10μg·g-1；弱抗旱苜蓿叶片相对含水量和SOD活性变化最敏感，且叶片相对含水量小于32.06%、SOD活性低于360.64 U·g-1；B) Under moderate stress of 0.18-0.33 g/mL, the leaves of strong drought-resistant alfalfa are most sensitive to changes in relative water content and hydrogen peroxide content, and the relative water content of leaves is greater than 50.43%, and the content of hydrogen peroxide is lower than 78.27 μmol·g -1 ; The content of reducing ascorbic acid in medium drought-resistant alfalfa is the most sensitive, and it is between 60.44-215.10 μg·g -1 ; The change of relative water content and SOD activity in leaves of weak drought-resistant alfalfa is the most sensitive, and the relative water content of leaves is less than 32.06%, and the SOD activity is low at 360.64 U·g -1 ;
C）0.33-0.43g/mL的重度胁迫，强抗旱苜蓿还原性抗坏血酸和还原性谷胱甘肽含量变化最敏感，且分别高于432.95μg·g-1和5280.17μmol·g-1；中抗旱苜蓿丙二醛（MDA）含量变化最敏感，且介于 0.13-0.15μmol·g-1；弱抗旱苜蓿过氧化氢酶活性、还原性谷胱甘肽含量、叶绿素含量和超氧阴离子自由基含量变化最敏感，且过氧化氢酶活性低于297.45 U·g-1·min-1、还原性谷胱甘肽含量低于2394.42μmol·g-1、叶绿素含量低于2.46 mg.g-1、超氧阴离子自由基含量高于152.92 nmol·g-1·min-1；C) Under the severe stress of 0.33-0.43g/mL, the contents of reduced ascorbic acid and reduced glutathione in alfalfa with strong drought resistance were the most sensitive, and they were higher than 432.95μg·g -1 and 5280.17μmol·g -1 respectively; moderate drought resistance The content of malondialdehyde (MDA) in alfalfa is the most sensitive to change, and it is between 0.13-0.15 μmol·g -1 ; the catalase activity, reduced glutathione content, chlorophyll content and superoxide anion free radical content in weak drought-resistant alfalfa The changes are most sensitive, and the catalase activity is lower than 297.45 U·g -1 ·min -1 , the reduced glutathione content is lower than 2394.42 μmol·g -1 , the chlorophyll content is lower than 2.46 mg.g -1 , The superoxide anion free radical content is higher than 152.92 nmol·g -1 ·min -1 ;
D）同时满足所述A）、B）、C）中，强抗旱苜蓿指标群的各指标阈值要求，则归为强抗旱苜蓿；同时满足所述A）、B）、C）中，中抗旱苜蓿指标群的各指标阈值要求，则归为中抗旱苜蓿；同时满足所述A）、B）、C）中，弱抗旱苜蓿指标群的各指标阈值要求，则归为弱抗旱苜蓿品种。D) Satisfying the threshold requirements of each indicator group of strong drought-resistant alfalfa in A), B), and C) at the same time, it is classified as strong drought-resistant alfalfa; meeting the above-mentioned A), B), and C) at the same time, moderate drought resistance Alfalfa varieties that meet the threshold requirements for each index of the alfalfa index group are classified as medium drought-resistant alfalfa varieties; those that meet the threshold requirements for each indicator group of weak drought-resistant alfalfa in A), B), and C) are classified as weak drought-resistant alfalfa varieties.
最后应说明的是：以上所述仅为本发明的优选实施例而已，并不用于限制本发明，尽管参照前述实施例对本发明进行了详细的说明，对于本领域的技术人员来说，其依然可以对前述各实施例所记载的技术方案进行修改，或者对其中部分技术特征进行等同替换。凡在本发明的精神和原则之内，所作的任何修改、等同替换、改进等，均应包含在本发明的保护范围之内。Finally, it should be noted that: the above is only a preferred embodiment of the present invention, and is not intended to limit the present invention. Although the present invention has been described in detail with reference to the foregoing embodiments, for those skilled in the art, it still The technical solutions recorded in the foregoing embodiments may be modified, or some technical features thereof may be equivalently replaced. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principles of the present invention shall be included within the protection scope of the present invention.
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